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ADK | Adenylate kinase

AS16 3155   | Clonality: Polyclonal  |  Host: Rabbit  |  Reactivity: E. coli

ADK | Adenylate kinase in the group Antibodies Other Species / Bacteria at Agrisera AB (Antibodies for research) (AS16 3155)



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Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Add review

Product Information

Immunogen

Protein derived from Escherichia coli, Uniprot: P69441

Host Rabbit
Clonality Polyclonal
Purity Serum
Format Lyophilized
Quantity 50 ĩl
Reconstitution For reconstitution add 50 ĩl, of sterile water
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications Western blot (WB)
Recommended dilution 1: 100 000 (WB)
Expected | apparent MW

23.5 kDa (E.coli), 24.3 kDa (S.cerevisiae)

Reactivity

Confirmed reactivity Escherichia coli
Predicted reactivity  
Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

Western blot using anti-ADK antibodies

Protein samples were prepared using TCA-extraction method. S. cerevisiae adk1Δ strains expressing either S. cerevisiae Adk1 protein (24.3 kDa) or E. coli adk protein (23.5 kDa) were cultivated logarithmically, and 5 OD-units of cells were collected for TCA-extraction method. In the end, protein pellets were re-suspended with 300 μl of TCA-Laemmli Loading Buffer and denatured at 100°C for 10 minutes. Total protein samples (8 μl of each sample) were separated by 12% SDS-PAGE and transferred to nitrocellulose membranes using a wet transfer protocol. The membranes were blocked with 5% dry-milk in 1x TBST for 1.5 hours at room temperature (RT) with agitation and then they were incubated with primary antibody (anti-adk antibody, 1:2,000) in 5% dry-milk in 1x TBST overnight at 4°C with agitation. The primary antibody solution was decanted and the membranes were washed 3 times for 10 minutes with 1x TBST with agitation. The membranes were then incubated with secondary antibody (HRP conjugated anti-rabbit IgG, 1:4,000) in 5% dry-milk in 1x TBST for 1 hour at RT with agitation. The membranes were washed as above and the signals were detected using chemiluminescent detection reagent and exposing the membranes in LAS4000 machine (GE-healthcare) for 10 seconds. Followed-up experiments revealed that lowering the primary antibody concentration from 1:2,000 to 1:50,000-100,000 reduces the unspecific background signals and lowers the risk of target signal oversaturation.

Courtesy of Dr. Hasan Tukenmez, Umeå University, Sweden

Additional information

Related products

Background

Background

Adenylate kinase (ADK) is important in cellular energy homeostasis. It is a phosphotransferase enzyme, catalyzing the interconversion of adenine nucleotides.

Alternative name: myokinase

Product citations

Selected references Tükenmez et al. (2016). Linkage between Fitness of Yeast Cells and Adenylate Kinase Catalysis. PLoS One. 2016 Sep 19;11(9):e0163115. doi: 10.1371/journal.pone.0163115. eCollection 2016.
immunogen:

Protein derived from Escherichia coli, Uniprot: P69441

Reconstitution: For reconstitution add 50 ĩl, of sterile water
Host: Rabbit
Clonality: Polyclonal
Purity: Serum
Format: Lyophilized
Quantity: 50 ĩl
storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications: Western blot (WB)
recommended dilution: 1: 100 000 (WB)
calculated | apparent molecular mass [kDa]:

23.5 kDa (E.coli), 24.3 kDa (S.cerevisiae)

Confirmed reactivity: Escherichia coli
predicted reactivity:  
not reactive in: No confirmed exceptions from predicted reactivity are currently known
Picture (footer): Application example

Western blot using anti-ADK antibodies

Protein samples were prepared using TCA-extraction method. S. cerevisiae adk1Δ strains expressing either S. cerevisiae Adk1 protein (24.3 kDa) or E. coli adk protein (23.5 kDa) were cultivated logarithmically, and 5 OD-units of cells were collected for TCA-extraction method. In the end, protein pellets were re-suspended with 300 μl of TCA-Laemmli Loading Buffer and denatured at 100°C for 10 minutes. Total protein samples (8 μl of each sample) were separated by 12% SDS-PAGE and transferred to nitrocellulose membranes using a wet transfer protocol. The membranes were blocked with 5% dry-milk in 1x TBST for 1.5 hours at room temperature (RT) with agitation and then they were incubated with primary antibody (anti-adk antibody, 1:2,000) in 5% dry-milk in 1x TBST overnight at 4°C with agitation. The primary antibody solution was decanted and the membranes were washed 3 times for 10 minutes with 1x TBST with agitation. The membranes were then incubated with secondary antibody (HRP conjugated anti-rabbit IgG, 1:4,000) in 5% dry-milk in 1x TBST for 1 hour at RT with agitation. The membranes were washed as above and the signals were detected using chemiluminescent detection reagent and exposing the membranes in LAS4000 machine (GE-healthcare) for 10 seconds. Followed-up experiments revealed that lowering the primary antibody concentration from 1:2,000 to 1:50,000-100,000 reduces the unspecific background signals and lowers the risk of target signal oversaturation.

Courtesy of Dr. Hasan Tukenmez, Umeå University, Sweden
background:

Adenylate kinase (ADK) is important in cellular energy homeostasis. It is a phosphotransferase enzyme, catalyzing the interconversion of adenine nucleotides.

Alternative name: myokinase

All references: Tükenmez et al. (2016). Linkage between Fitness of Yeast Cells and Adenylate Kinase Catalysis. PLoS One. 2016 Sep 19;11(9):e0163115. doi: 10.1371/journal.pone.0163115. eCollection 2016.

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